In vitro anthelmintic activity of Brassica nigra Linn ... - URP Journals

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International Journal of Natural Products Research
Universal Research Publications. All rights reserved
Original Article
In vitro anthelmintic activity of Brassica nigra Linn. seeds
Abstract
Nitinkumar Upwar*, Roshan Patel, Naheed Waseem 1 and Naveen Kumar Mahobia 2
Department of Pharmacognosy, Shree Leuva Patel Trust Pharmacy Mahila College, Amreli (Gujarat)
1 Department of Pharmacology, Shree Leuva Patel Trust Pharmacy Mahila College, Amreli (Gujarat)
2 Department of Pharmaceutical Chemistry, Shree Leuva Patel Trust Pharmacy Mahila College, Amreli (Gujarat)
E-mail*: nitinupwar@yahoo.com
Received 17 March 2011; accepted 26 March 2011
Alcohoilc extract from the seeds of Brassica nigra Linn. were investigated for their anthelmintic activity against Pheretima
posthuma and Ascardia galli. Various concentrations (10-100 mg/ml) of extract were tested in bioassay, which involved
determination of time of paralysis and time of death of the worms. Alcoholic extracts exhibited significant activity at highest
concentration of 100mg/ml. Piperazine citrate (10mg/ml) was included as standard reference and distilled as control.
© 2011 Universal Research Publications. All rights reserved
Key words: Brassica nigra Linn, anthelmintic activity, Pheretima posthuma and Ascardia galli
Introduction
Brassica nigra Linn. (Brassicaceae) is commonly known as
‘Mustard’ in English and ‘Sarson’ in Hindi. The plant is
cultivated throughout India and central Europe. It is an erect,
simple, mostly branched, annual herb, 50 to 60 cm tall with
lower leaves distinctly stalked, lyrate-pinnatisect, largest and
ovate middle leaves shorter-petioled and upper leaves
oblong-linear mostly entire and all very bright green.
Racemose inflorescence, fruit are long erect pods, seeds
globose, 1mm. diameter, obscurely brown and black near the
hilum, delicately alveolate. The extract of Brassica nigra
Linn was reported as antidiabetic and anthperlipidemic effect.
The leaves hot with a pleasant taste increase the bile,
vermicide, good for throat complaints and seeds are bitter,
hot and acrid used to cure enlargement of the spleen and
dispel fever, increase the bile, remove cough, tumors,
anthelmintic, increase appetite, cure skin diseases, itching and
destroy external parasites is mentioned in Ayurveda. Poultice
of seed are used counter irritant in several complaints of
nervous system, pneumonia and an emetic in narcotic
poisoning. The seeds contain alkaloid, sinapine, myrosin,
sinigrin, inosite, albumins, gums and colouring matters [1]. In
spite of its traditional use as anthelmintic, there are no reports
on systematic and scientific study of anthelmintic activity of
Brassica nigra in the present study; we investigate the
anthelmintic activity of alcoholic extracts of seeds of
Brassica nigra Linn.
Materials and Methods
Plant material
The seeds of Brassica nigra Linn. was purchased from
locally were identified and authenticated by Department of
Botany, Smt. U. B. Bhagat Science Mahila college, Amreli,
Gujarat. The powdered seed was extracted with methanol by
Soxahlet apparatus. The solvent was concentrated
byevaporating methanol using a rotary evaporator.
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Table 1: Anthelmintic activity of alcoholic extract of Brassica nigra Linn.
Time taken for paralysis (P) and death (D) of worms (mins)
Concentration
Test
P. posthuma A. galli
(mg/ml)
P D P D
Control - - - - -
Alcohol
extracts
Piperazine
citrate
10 24.40 ± 0.4 64.17 ± 0.5 16.50 ± 0.5 43.54 ± 0.5
50 17.26 ± 0.1 44.64 ± 0.4 10.51 ± 0.7 24.61 ± 0.4
100 8.15 ± 0.2 27.45 ± 0.3 7.94 ± 0.4 16.09 ± 0.8
10 20.37 ± 0.3 51.35± 0.1 12.05 ± 0.91 40.41 ± 0.3
Results are expressed as Mean ± SEM from six observations
Preparation of extracts
The seeds of Brassica nigra Linn. cleaned and coarsely
powder. It was then passed through the 40 mesh sieve. A
weighted quantity (200 gm) of the power was subjected to
continuous hot extraction in Soxhlet Apparatus exhaustively.
The extract was evaporated under pressure using rotary
evaporator until all solvent has been removed to give an
extract sample. Percentage yield of methanol extract 4.3%
w/w. The dry extracts obtained were subjected to various
chemical tests to detect the presence of different
phytoconstituents [2,3].
Animals
Indian adult earth worm (Pheretima posthuma) was collected
from water logged areas of Amreli and Ascardia galli
(nematode) worm were obtained from freshly slaughtered
fowls. Both worm types were identified at the Department of
Zoology, Smt. U. B. Bhagat Science Mahila college, Amreli,
Gujarat, India.
Drugs and chemicals used
Piperazine citrate (Noel, Mumbai) were used as reference
standards. Chemicals: Methanol (95% v/v) (Rexol Ecofuels
P. Ltd. Mumbai).
Evaluation of anthelmintic activity
The assay was performed on Indian earthworm Pheritima
posthuma due to its anatomical and physiological
resemblance with intestinal roundworm parasite of human
beings [4-8]. Because of easy availability, has been used
widely for the initial evaluation anthelmintic compounds in
vitro [9-13]. Ascardia galli worms are easily available in
plenty from freshly slaughtered flows and their use, as
suitable model for screening of anthelmintic drug was
advocated as earlier [14-16]. Six groups of worms were used
to assess the anthelmintic properties of alcoholic extracts of
Brassica nigra Linn. seeds. Groups 1 were the control worms
placed in distilled water; groups 2-4 were treated with 10, 50
and 100 mg/ml of methanolic extracts Brassica nigra Linn. In
distilled water; group 5 with Piperazine citrate (10mg/ml) in
distilled water. Each group included six worms of each type.
Observations were made for the time taken to set paralysis
and death of the individual worms. Mean time for the
paralysis (P) in min. was noted when no movement of any
sort could be observed, except when the worm was shaken
vigorously; time of death (D) in min. was recorded after
ascertaining the worms neither moved when shaken
vigorously nor when dipped in warm water (50°C).
Piperazine citrate (10 mg/ml) was included as reference
compound [17].
Results
Preliminary phytochemical screening of alcoholic extract
revealed presence of glycosides and alkaloids. As shown in
Table 1, the alcoholic extract of seeds of Brassica nigra Linn.
Displayed significant anthelmintic properties at higher
concentration. Alcoholic extract showed anthelmintic
activities in dose dependant manner giving shortest of time of
paralysis (P) and death (D) with 100 mg/ml concentration for
both type worms. The alcoholic extract of seeds of Brassica
nigra Linn. Caused paralysis in 8.15 min and death in 27.45
min against the earthworm Pheretima posthuma. The
reference drug piperazine citrate showed same at 21 min and
59 min.
Ascardia galli worms also showed sensitivity to the alcoholic
extracts of Brassica nigra Linn. The alcoholic extract caused
paralysis in 5 min, and death in 29 min at higher
concentration of 100mg/ml. Piperazine citrate did same at 12
and 41min.
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Discussion
In this study, anthelmintic assay was performed on adult
Indian earthworm, Pheretima posthuma due to its anatomical
and physiological resemblance with the intestinal roundworm
parasite of human beings [5-8]. Because of easy availability,
earthworms have been used widely for the initial evaluation
of anthelmintic compounds in vitro [18-19]. Ascardia galli
worms are easily available in plenty from freshly slaughtered
fowls and their use, as a suitable model for screening of
anthelmintic drug was advocated earlier [14-16]. Piperazine
citrate causes flaccid paralysis of worms that resulting
expulsion of worms by peristalsis. Piperazine citrate has
causes death of the parasite. Therefore, it is concluded that
alcoholic extracts of Brassica nigra Linn. Seeds have potent
anthelmintic activity when compared the conventionally used
drug and is equipotent to standard anthelmintic drug tested
against worm species. It justifies its Ayurvedic use in curing
helmintic infections.
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Source of support: Nil; Conflict of interest: None declared
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